Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging

Stefanie Kuhns, Alice Dupont Juhl, Zeinab Anvarian, Daniel Wüstner, Lotte B. Pedersen, Jens S. Andersen*


Publikation: Kapitel i bog/rapport/konference-proceedingKapitel i bogForskningpeer review


CRISPR-mediated endogenous tagging of genes provides unique possibilities to explore the function and dynamic subcellular localization of proteins in living cells. Here, we describe experimental strategies for endogenous PCR-tagging of ciliary genes in human RPE1 cells and how image acquisition and analysis of the expressed fluorescently tagged proteins can be utilized to study the dynamic ciliary processes of intraflagellar transport and vesicular trafficking.

TitelCilia : Methods and Protocols
RedaktørerVito Mennella
ForlagHumana Press
ISBN (Trykt)978-1-0716-3509-4, 978-1-0716-3506-3
ISBN (Elektronisk)978-1-0716-3507-0
StatusUdgivet - 2024
NavnMethods in Molecular Biology

Bibliografisk note

Funding Information:
We thank Joel Pomerantz, Kazuhisa Nakayama, Michael Knop, Keith Joung, and Benjamin Kleinstiver for reagents and the Danish Molecular Biomedical Imaging Center, University of Southern Denmark, for use of imaging equipment, supported by Novo Nordisk Foundation (NNF18SA0032928). This work was supported by grants from Independent Research Fund Denmark (grant 8021-00425B) to J.S.A. and Novo Nordisk Foundation (NNF18OC0053024, NNF15OC0016886 and NNF14OC0011535), The Danish Cancer Society (R146-A9590) and Hartmann Foundation (A31662) to L.B.P.


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